The recent technological advances in top-down silicon nanotechnology and micromachining of semiconductor materials present themselves with new opportunities for small, sensitive, one time use, point of care diagnostic devices capable of rapid and highly accurate analysis of samples of body fluid. Nano- sensors and microfluidic biochips have been successfully demonstrated for the detection of proteins in fluids, yet still to be realized in a robust array format, with detection of multiple proteins, which can be used in preclinical studies or clinical samples. In this proposal, we have assembled a truly comprehensive team of interdisciplinary researchers with the goal of applied and translational multidisciplinary research for designing and producing robust top-down silicon-based field-effect nano-sensor platform technologies integrated in one-time-use point-of-care diagnostic biochips, functionalized with multiple antibodies, for the ex-vivo detection of cancer proteins from cell lysate from breast aspirates. These tasks will be accomplished by our team of researchers with expertise in clinical translation (Susan Clare at ILJSOM), micro/nano-fabrication of silicon based field effect devices and on-chip sample concentration and separation (Rashid Bashir at Purdue and Luke Lee at UC Berkeley), Simulation and computation of nanosensors (Ashraf Alam at Purdue), and Cancer bio-chemistry (Don Bergstrom at Purdue). The following are the specific aims of the project: (i) Develop on chip cell lysing approaches from breast aspirates, (ii) Develop novel techniques to functionalize the sensor array surfaces with antibodies, while minimizing non-specific adsorption and bio-fouling, (iii) Develop novel computational and simulation strategies for modeling the field effect sensor response upon protein binding in fluids, (iv) Develop robust top-down silicon-based field-effect nano-plate arrays for multiplexed detection of cancer proteins & markers, and (v) Develop integrated biochip sensors and perform extensive tests and preclinical studies. We will keep the focus on the issues and requirements towards assay development to perform ex-vivo preclinical studies with human samples.